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抗植物血凝素(PHA)的中国仓鼠卵巢细胞突变体在与含有PHA受体糖蛋白的脂质体融合后,会恢复对这种凝集素的敏感性。

Phytohemagglutinin (PHA)-resistant Chinese hamster ovary cell mutants acquire sensitivity to the lectin after fusion with liposomes containing PHA receptor glycoproteins.

作者信息

Letellier M, Dupuis G

出版信息

Biochem Biophys Res Commun. 1985 Nov 15;132(3):1024-30. doi: 10.1016/0006-291x(85)91909-6.

Abstract

Phytohemagglutinin receptor glycoproteins have been inserted into phospholipid vesicles and these have been fused with phytohemagglutinin-resistant chinese hamster ovary cells. Our results show that the fused cells acquire "neoreceptors" for the lectin phytohemagglutinin. Fluorescence activated cell sorting analyses show that approximately 40% of the cells fused with the receptor-containing vesicles. Studies with 125I-labelled lectin showed that fused cells bound three times more ligand than untreated mutant cells. Furthermore, lectin receptors were functionally inserted in the mutant cell plasma membrane. Fused cells cultured in the presence of lectin (200 micrograms ml-1) lost rapidly (8 hours or less) their ability to incorporate [3H] thymidine. Whereas mutant cells cultured for 16 hours in the presence of 50-400 micrograms ml-1 of lectin remained viable, fused cells showed a 45% decrease in 3H-labelled nucleotide incorporation. The method described here should be of general applicability for the study of lectin-dependent cytotoxicity in chinese hamster ovary cell lines.

摘要

植物血凝素受体糖蛋白已被插入磷脂囊泡中,并且这些囊泡已与抗植物血凝素的中国仓鼠卵巢细胞融合。我们的结果表明,融合细胞获得了针对凝集素植物血凝素的“新受体”。荧光激活细胞分选分析表明,约40%的细胞与含受体的囊泡融合。用125I标记的凝集素进行的研究表明,融合细胞结合的配体比未处理的突变细胞多三倍。此外,凝集素受体在功能上被插入突变细胞质膜中。在凝集素(200微克/毫升)存在下培养的融合细胞迅速(8小时或更短时间)丧失掺入[3H]胸苷的能力。而在50 - 400微克/毫升凝集素存在下培养16小时的突变细胞仍保持活力,融合细胞的3H标记核苷酸掺入量减少了45%。这里描述的方法对于研究中国仓鼠卵巢细胞系中凝集素依赖性细胞毒性应具有普遍适用性。

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